TaRBP1 stabilizes TaGLTP and negatively regulates stripe rust resistance in wheat

The dynamic balance and distribution of sphingolipid metabolites modulate the level of programmed cell death and plant defence. However, current knowledge is still limited regarding the molecular mechanism underlying the relationship between sphingolipid metabolism and plant defence. In this study, we identified a wheat RNA-binding protein 1 (TaRBP1) and TaRBP1 mRNA accumulation significantly decreased in wheat after infection by Puccinia striiformis f. sp. tritici (Pst). Knockdown of TaRBP1 via virus-induced gene silencing conferred strong resistance to Pst by enhancing host plant reactive oxygen species (ROS) accumulation and cell death, indicating that TaRBP1 may act as a negative regulator in response to Pst. TaRBP1 formed a homopolymer and interacted with TaRBP1 C-terminus in plants. Additionally, TaRBP1 physically interacted with TaGLTP, a sphingosine transfer protein. Knockdown of TaGLTP enhanced wheat resistance to the virulent Pst CYR31. Sphingolipid metabolites showed a significant accumulation in TaGLTP-silenced wheat and TaRBP1-silenced wheat, respectively. In the presence of the TaRBP1 protein, TaGLTP failed to be degraded in a 26S proteasome-dependent manner in plants. Our results reveal a novel susceptible mechanism by which a plant fine-tunes its defence responses by stabilizing TaGLTP accumulation to suppress ROS and sphingolipid accumulation during Pst infection.     

Microscopic Observation on the Development of Puccinia striiformis in the Spike and Stem of Wheat

The majority of germ tubes of the pathotype CYR32 of Puccinia striiformis f.sp. tritici formed on the surface of spike organs of the susceptible wheat cv. Suwon 11 penetrated through the stomatal pore, only a few germ tubes formed small appressoria over the stomata. In the lemma, palea and glume, the stripe rust fungus spread between the parenchyma cells close to the inner epidermal layer, but the fungus did not develop between the thick‐walled cells near the outer epidermal layer of these organs. In the awn and stem, spread of the stripe rust was confined to the intercellular spaces of the chlorophyll parenchyma, beneath the invaded stomatal pore of the epidermis and the urediniospores to be released disrupted the epidermis. In the caryopsis, the spread of hyphae was restricted to the intercellular spaces of the pericarp cells.     

Atmospheric teleconnection patterns associated with wheat stripe rust disease in North China

 Previous work has shown that long-term (>40 years) time series of wheat stripe rust disease from North China contain a signal related to the Southern Oscillation Index (SOI). However, no cause-and-effect relationships or direct links between SOI and disease have been established. Because mid-latitude teleconnection patterns form important links between global atmospheric variations and regional weather anomalies, such as the Southern Oscillation, studies of such patterns could result in a better physical and biological interpretation for the SOI-disease association observed previously. We used cross-spectral analyses to determine if and on which time scales severity of stripe rust in five regions of North China between 1952 and 1990 was associated with fluctuations in four Northern Hemisphere teleconnection patterns. These included the Pacific/North American, the Western Atlantic, the Western Pacific, and the Eurasian patterns. The analysis showed consistent and significant (P≤0.10) coherence relationships between the Western Atlantic (WA) pattern and stripe rust severity at a periodicity of 3.00 years. The phase relationships showed that, at the 3.00-year periodicity, the WA series and the disease series were out of phase by about half a period in all five regions. This phase difference indicated that peaks in the WA series, which are associated with below-normal winter temperatures over much of China, coincided with troughs in the disease series (i.e. low disease severity), presumably because of the negative effect of low winter temperatures on survival of the stripe rust pathogen. The analysis further showed that the WA series and the SOI series were highly significantly (P≤0.01) coherent at a periodicity of 4.33 years, which could explain the association between SOI and stripe rust severity reported previously. Received: 12 February 1997 / Accepted: 18 May 1998     

Study of Genetic Variability Among Monopycnidial and Monopycnidiospore Isolates Derived from Single Pycnidia of Stagonospora ssp. and Septoria tritici with the use of RAPD-PCR, MP-PCR and rep-PCR Techniques

Thirty-six isolates of Stagonospora avenae f. sp. triticea, S. nodorum and Septoria tritici recovered from asexual fruiting bodies - pycnidia and their spores were assessed for DNA polymorphism with the use of three molecular techniques: microsatellite-primed polymerase chain reaction (MP-PCR), primers correspond to dispersed repetitive elements (rep-PCR) and random amplified polymorphic DNA (RAPD-PCR). These polymerase chain reaction (PCR)-based techniques were simultaneously evaluated for their capacity to detect genetic variation at DNA level. The most polymorphic DNA profiles of monopycnidial and monopycnidiospore isolates were detected with diverse microsatellite motifs used for PCR priming. The lowest similarity values 0.86, 0.76 and 0.84 were identified among monopycnidiospore isolates derived from the same pycnidium of S. avenae f. sp. triticea, S. nodorum and S. tritici, respectively. The above, rather low similarities, found for isolates recovered from single pycnidia, supported a hypothesis that heterokaryosis resulted from high mutation rate of microsatellites and transposons activity. This would have fundamental consequences for the genetic status of asexual populations of Stagonospora spp. and S. tritici. The data produced by this study indicate that more attention should be paid to asexual reproduction as a possible source of genetic variability among populations of the pathogens.     

感染叶锈菌的小麦细胞间隙液中激发子的定性及初步分离

感染叶锈菌后小麦叶片细胞间隙液(1wF)中有激发子存在,它(们)能诱导寄主PAL、PO活性的增强及细胞过敏性坏死的产生。这种有诱导活性的物质分别用NaIO4和蛋白酶处理证明含有糖基和蛋白质成分,可能是糖蛋白。IWF经凝胶过滤分离后,各部分诱导不同的防卫反应的活性强度不等。因此IWF中可能含有几种不同成分的激发子,或是同种成分而聚合度不同的激发子。     

Development of simple sequence repeat markers for the plant pathogenic rust fungus,Puccinia graminis

Twenty‐four dinucleotide simple sequence repeat markers were developed for the phytopathogenic fungus, Puccinia graminis. The identified loci were polymorphic, with allelic diversity ranging from two to 11 alleles. Observed and expected levels of heterozygosity ranged from 0.000 to 0.960 and from 0.113 to 0.846, respectively. Fourteen of the loci deviated significantly from Hardy–Weinberg equilibrium. Null alleles were observed for 10 of the 24 loci with a frequency of 4–16%. A preliminary screen of other Puccinia cereal rust fungi (P. coronata, P. striiformis and P. triticina) indicated that these primer pairs are specific to P. graminis.     

Single residues in the LRR domain of the wheat PM3A immune receptor can control the strength and the spectrum of the immune response

The development of improved plant nucleotide‐binding, leucine‐rich repeat (LRR) immune receptors (NLRs) has mostly been based on random mutagenesis or on structural information available for specific receptors complexed with the recognized pathogen effector. Here, we use a targeted mutagenesis approach based on the natural diversity of the Pm3 powdery mildew resistance alleles present in different wheat (Triticum aestivum) genotypes. In order to understand the functional importance of the amino acid polymorphisms between the active immune receptor PM3A and the inactive ancestral variant PM3CS, we exchanged polymorphic regions and residues in the LRR domain of PM3A with the corresponding segments of PM3CS. These novel variants were functionally tested for recognition of the corresponding AVRPM3^A2/F2 avirulence protein in Nicotiana benthamiana. We identified polymorphic residues in four regions of PM3A that enhance the immune response, but also residues that reduce it or result in complete loss of function. We found that the identified critical residues in PM3A modify its activation threshold towards different protein variants of AVRPM3^A2/F2. PM3A variants with a lowered threshold gave a stronger overall response and gained an extended recognition spectrum. One of these variant proteins with a single amino acid change was stably transformed into wheat, where it conferred race‐specific resistance to mildew. This is a proof of concept that improved PM3A variants with an enlarged recognition spectrum can be engineered based on natural diversity by exchanging single or multiple residues that modulate resistance function.     

Thermal adaptation in the fungal pathogen Mycosphaerella graminicola

Genetic differentiation in thermal adaptation can result from a trade-off between the performance of organisms across different temperatures or from the accumulation of deleterious mutations. In this experiment, we assayed thermal sensitivity of 138 genetically distinct Mycosphaerella graminicola isolates sampled from five host populations in four locations under two temperature regimes (22 and 15 °C) and found significant differences in growth rate and response to temperature among populations. On average, genetic differentiation accounted for more than 50% of phenotypic variation in thermal adaptation while plasticity contributed less than a quarter of phenotypic variation. Populations originating from warm places performed better under the high-temperature regime and had a larger positive response to increasing temperature. Pairwise population differentiation (Q(ST) ) in temperature sensitivity, measured by taking the ratio of growth rates at 22 to 15 °C, was positively and significantly correlated to the pairwise difference in annual mean temperature at the collection sites. Because overall Q(ST) in temperature sensitivity was significantly higher than overall G(ST) in neutral restriction fragment length polymorphism loci, we believe that the primary mechanism underlying this thermal adaptation is antagonistic pleiotropy. Our results indicate that temperature sensitivity is a better indicator of thermal adaptation than growth rate at individual temperatures.     

Pathotypes of Blumeria graminis f. sp. tritici,the causal agent of wheat powdery mildew from some regions of Iran

Wheat powdery mildew is controlled mainly by race-specific resistance. To be effective, breeding wheat for resistance to powdery mildew requires knowledge of virulence diversity in local populations of the pathogen. Isolates of Blumeria graminis, collected in 2009 and 2010 from three areas of Iranian production, were analysed for virulence using a host differential series comprised of 16 known genes conferring resistance to powdery mildew. The results showed that high-virulence frequencies to genes Pm1, Pm2, Pm4a, Pm5, Pm6, Pm7, Pm8 and Pm9 were found over both years and across all three areas. Virulence frequencies for Pm3a and Pm3b were intermediate, while virulence frequencies for Pm3a, Pm3c, Pm4a and Pm2, 6 were low. Genes Pm1, 2, 9 and Pm2, 4b, 8 were highly resistant in all regions. Virulence to Pm8 increased to high levels, while virulence to Pm4a decreased across the area surveyed from 2009 to 2010.